Generation of Organoids from Mouse Extrahepatic Bile Ducts

Junya Shiota; Nureen  H. Mohamad Zaki; Juanita  L. Merchant; Linda  C. Samuelson; Nataliya Razumilava

doi:10.3791/59544

JoVE Journal
Developmental Biology

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JoVE Journal Developmental Biology

Generation of Organoids from Mouse Extrahepatic Bile Ducts

Organoids 마우스 Extrahepatic 담 즙 덕트에서의 세대

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09:13 min

April 23, 2019

DOI: 10.3791/59544-v

Junya Shiota¹, Nureen H. Mohamad Zaki¹, Juanita L. Merchant^1,2, Linda C. Samuelson^1,2, Nataliya Razumilava¹

¹Departments of Internal Medicine,The University of Michigan, ²Molecular & Integrative Physiology,The University of Michigan

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Overview

This protocol describes the production of a mouse extrahepatic bile duct 3-dimensional organoid system. These biliary organoids can be maintained in culture to study cholangiocyte biology and are composed of polarized epithelial cells.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Organ Models

Background

Limited access to preclinical cholangiopathy models.
Challenges with pluripotent stem cell and liver-derived cholangiocyte organoid models.
Need for adult-tissue specific and reproducible models.
Importance of studying cholangiocyte biology.

Purpose of Study

To develop a model for studying cholangiocyte biology.
To provide a reductionist and cost-efficient method for organoid culture.
To facilitate research in laboratories lacking access to human tissues.

Methods Used

Isolation of intrahepatic bile duct from adult mice.
Maintaining organoids in culture.
Studying tissue regeneration and cell-to-cell interactions.
Demonstration of the procedure by a post-doc researcher.

Main Results

Successful production of biliary organoids.
Organoids express markers of progenitor and biliary cells.
Capability of culturing an unlimited number of cholangiocytes.
Potential for studying cholangiocyte biology and interactions.

Conclusions

The developed organoid system is a valuable tool for cholangiocyte research.
It addresses limitations of existing models.
Offers a reproducible method for studying cholangiopathies.

Frequently Asked Questions

What are biliary organoids?

Biliary organoids are 3D structures derived from the extrahepatic bile duct that can be cultured to study cholangiocyte biology.

Why is this model important?

It provides a specific and reproducible model for studying cholangiopathies, especially for labs without access to human tissues.

How are the organoids maintained?

The organoids can be maintained in culture, allowing for extensive study of their properties and behaviors.

Who demonstrated the procedure?

The procedure was demonstrated by Junya Shiota, a post-doc from the laboratory.

What are the advantages of this model?

The model is adult-tissue specific, reductionist, reproducible, and time and cost efficient.

What can be studied using these organoids?

Researchers can study tissue regeneration and cell-to-cell interactions in cholangiocytes.

이 프로토콜 마우스 extrahepatic 담 즙 덕트 3 차원 organoid 시스템의 생산을 설명합니다. 이러한 담 즙 organoids cholangiocyte 생물학 연구 문화에서 유지할 수 있습니다. 담 즙 organoids 조상 및 담 즙 셀의 마커를 표현 하 고 편광 된 상피 세포의 구성 됩니다.

이 뮤린 외형 담관 오르가노이드 시스템은 전임상 담관증 모델에 대한 제한된 접근과 다능성 줄기 세포 및 간 유래 콜린고이티테 오르가노이드 모델의 한계를 해결할 수 있습니다. 우리의 모델은 성인 조직 특정, 감소주의자, 재현 가능, 시간과 비용 효율적인. 그것은 인간의 조직에 액세스 할 수없는 실험실에 특별한 도움이 될 것입니다.

우리의 기술은 조직 재생 및 세포 대 세포 상호 작용을 연구하기 위하여 외형 담관 cholangiocytes의 거의 무제한 수의 배양을 허용합니다. 절차를 시연하는 것은 내 실험실에서 박사 후 의사 인 주야 시오타 (Junya Shiota)가 될 것입니다. 간 내 담관 격리의 경우 성인 마우스를 척추 위치에 놓고 중간선을 따라 복강을 엽니다.

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